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用一种简单技术切割低浓度聚丙烯酰胺凝胶对核糖体和病毒核糖核酸进行电泳分析。

Electrophoretic analysis of ribosomal and viral ribonucleic acids with a simple technique for slicing low-concentration polyacrylamide gels.

作者信息

Schaffer F L, Soergel M E, Straube D C

出版信息

Appl Microbiol. 1971 Oct;22(4):538-45. doi: 10.1128/am.22.4.538-545.1971.

Abstract

The electrophoretic mobilities of ribosomal ribonucleic acids (RNA) from cultured mammalian (HeLa, Vero, MDBK), avian (chick embryo), and bacterial (Escherichia coli) cells, and RNA species extracted from selected viruses (Sindbis, polio, tobacco mosaic, Sendai) were compared, employing a simple, inexpensive technique for slicing low-concentration polyacrylamide gels. The procedure provides for rapid fractionation of gels used for characterization of RNA, incorporating extrusion and serial sectioning of frozen gels. Among 28S ribosomal RNA species, Vero and MDBK were indistinguishable, whereas HeLA RNA had a slightly lower mobility (higher apparent molecular weight) and chick RNA had a higher mobility (lower apparent molecular weight). The 18S ribosomal RNA species of the three mammalian sources were indistinguishable, but chick 18S RNA had a slightly lower apparent molecular weight. The inverse relation between mobility and log-molecular weight among the ribosomal and viral RNA species, though not highly precise, demonstrates the applicability of the technique to the study of molecular weights of viral RNA species.

摘要

采用一种简单、廉价的低浓度聚丙烯酰胺凝胶切片技术,比较了培养的哺乳动物(海拉细胞、非洲绿猴肾细胞、牛肾细胞)、禽类(鸡胚)和细菌(大肠杆菌)细胞的核糖体核糖核酸(RNA)以及从选定病毒(辛德毕斯病毒、脊髓灰质炎病毒、烟草花叶病毒、仙台病毒)中提取的RNA种类的电泳迁移率。该方法通过对冷冻凝胶进行挤压和连续切片,实现了用于RNA表征的凝胶的快速分级分离。在28S核糖体RNA种类中,非洲绿猴肾细胞和牛肾细胞的RNA无法区分,而海拉细胞的RNA迁移率略低(表观分子量较高),鸡胚RNA的迁移率较高(表观分子量较低)。三种哺乳动物来源的18S核糖体RNA种类无法区分,但鸡胚18S RNA的表观分子量略低。核糖体和病毒RNA种类的迁移率与对数分子量之间的反比关系虽然不太精确,但证明了该技术在研究病毒RNA种类分子量方面的适用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8248/376358/2bbf1e2009e7/applmicro00120-0077-a.jpg

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